Background Although several cell culture systems have been developed to investigate the function of the mam-mary gland in dairy livestock,they have potential limitations,such as the loss of alveolar structure or genet...Background Although several cell culture systems have been developed to investigate the function of the mam-mary gland in dairy livestock,they have potential limitations,such as the loss of alveolar structure or genetic and phe-notypic differences from their native counterparts.Overcoming these challenges is crucial for lactation research.Development of protocols to establish lactating organoid of livestock represents a promising goal for the future.In this study,we developed a protocol to establish a culture system for mammary organoids in dairy goats to model the mammary gland development and lactation process.Results The organoids cultured within an extracellular matrix gel maintained a bilayer structure that closely resem-bled the native architecture of mammary tissue.The expansion of mammary organoids was significantly promoted by growth factors containing epidermal growth factor and fibroblast growth factor 2 whereas the proliferative index of the organoids was significantly inhibited by the treatment with WNT inhibitors.Upon stimulation with a lactogenic medium containing prolactin,the mammary organoids exhibited efficient lactation,characterized by the accumu-lation of lipid droplets in the lumen space.The lactation could be sustained for more than 3 weeks.Importantly,the expression patterns of genes related to fatty acid synthesis and milk proteins in lactating organoids closely mirrored those observed in mammary tissues.These observations were confirmed by data from proteomic analysis that the bulk of milk proteins was produced in the lactating organoids.Conclusion This study is the first to establish a mammary organoid culture system modeling the mammary gland development and lactation process in ruminants.The efficient induction of lactation in ruminant mammary organoids holds promises for advancing the field of cell-based milk bio-manufacture in the food industry.展开更多
基金Innovation 2030-Biological Breeding-Major Projects(2022ZD04014)Key R&D Program of Zhejiang Province(2022C04017).
文摘Background Although several cell culture systems have been developed to investigate the function of the mam-mary gland in dairy livestock,they have potential limitations,such as the loss of alveolar structure or genetic and phe-notypic differences from their native counterparts.Overcoming these challenges is crucial for lactation research.Development of protocols to establish lactating organoid of livestock represents a promising goal for the future.In this study,we developed a protocol to establish a culture system for mammary organoids in dairy goats to model the mammary gland development and lactation process.Results The organoids cultured within an extracellular matrix gel maintained a bilayer structure that closely resem-bled the native architecture of mammary tissue.The expansion of mammary organoids was significantly promoted by growth factors containing epidermal growth factor and fibroblast growth factor 2 whereas the proliferative index of the organoids was significantly inhibited by the treatment with WNT inhibitors.Upon stimulation with a lactogenic medium containing prolactin,the mammary organoids exhibited efficient lactation,characterized by the accumu-lation of lipid droplets in the lumen space.The lactation could be sustained for more than 3 weeks.Importantly,the expression patterns of genes related to fatty acid synthesis and milk proteins in lactating organoids closely mirrored those observed in mammary tissues.These observations were confirmed by data from proteomic analysis that the bulk of milk proteins was produced in the lactating organoids.Conclusion This study is the first to establish a mammary organoid culture system modeling the mammary gland development and lactation process in ruminants.The efficient induction of lactation in ruminant mammary organoids holds promises for advancing the field of cell-based milk bio-manufacture in the food industry.
